Academic Sciences Asian Journal of Pharmaceutical and Clinical Research

Vol 5, Suppl 3, 2012 ISSN - 0974-2441

Research Article

Vol. 4, Issue 3, 2011

EVALUATION OF WOUND HEALING POTENTIAL OF CYNODON DACTYLON
ISSN - 0974-2441
PAYAL DANDE*, ANIS KHAN
SVKM’s NMIMS, SPTM, Shirpur, Dhule- 425 405, Maharashtra, India. Email: payal4nmims@yahoo.com
Received:23 May 2012, Revised and Accepted:21 June 2012
ABSTRACT
Objective: To evaluate the wound healing potential of Cynodon dactylon.
Study design: Ethnomedicinal claim strongly suggests the use of Cynodon dactylon L. as first aid in minor injuries in traditional medicine 10. To
evaluate its action, aqueous and alcoholic extract of Cynodon dactylon L. were prepared. Preliminary phytochemical studies were performed. Wound
healing potential was evaluated in different experimental model such as Excision wound healing model and Incision wound healing model in albino
wistar rats by using the gel preparation of aqueous and alcoholic extract.
Results: Preliminary phytochemical studies revealed the presence of carbohydrates, tannins, phenols, flavonoids, amino acid, proteins, alkaloids and
glycosides in both aqueous extract and alcoholic extract. Further, alcoholic extract & aqueous extract gel showed significant increased in the rate of
wound healing in excision model (p<0.05) and in the incision wound healing (p<0.01), indicating the wound healing potential of Cynodon dactylon.
Conclusion: The present study revealed that the aqueous and alcoholic extract of Cynodon dactylon L. has a significant wound healing potential and
supports its traditional claim to be used in burns & inflammation.
Keywords: Cynodon dactylon, excision wound healing model, incision wound healing model.
INTRODUCTION
Interruption in the cellular and anatomic continuity of a tissue leads Plant extraction
to formation of wound1. Wound can be caused due to various factors
such as chemical, physical, thermal, microbial or immunological Aqueous extract
harm. Such conditions lead to discomfort and are more prone to
infection and other troublesome complications2. Wound healing The aqueous extract of the herb was prepared by soaking 250 gm of
encompasses a series of steps that may ultimately lead to restoration herb in 2000 ml of distill water. It was further extracted in soxhlet
of total integrity of the damaged tissue. The current drugs available apparatus for 12 hrs at boiling temperature. The resulting extract
are associated with challenges of being expensive, causing allergy or was filtered and concentrated in equitron rotavapour under reduced
leading to drug resistance. To overcome these problems, researchers pressure. The concentrated extract was lyophilized to get a powder
are seeking the help of traditional herbs for getting a better (yield 2.8%, w/w).
alternative3. This has lead to the newer research studies that are
being carried to find new drugs which will not only promise fast Alcoholic extract
healing but also will reduce the complication and cost. The ayurvedic
The fresh plant upto 250 gm was cleaned with distilled water and
traditional practitioners from India have many such natural drugs
for treating wounds and burns. was successfully extracted with 70:30 w/v (ethanol: water) using
soxhlet apparatus. The extraction was carried out for 24 hrs at room
Cynodon dactylon Linn. is a member of the family Graminae temperature. The resulting extract was filtered and concentrated at
(Poaceae). It is a creeping grass, very tough, drought resistant, light 45oC in rotavapour under reduced pressure. The concentrated
green in color, has a coarse texture, and fast growing. It is found in extract was lyophilized to get a powder (yield 3.2% w/w).
short cylindrical pieces about 3 to 20 mm long & 2 to 3 or sometimes
4 mm in diameter5. Cynodon dactylon aqueous extract have been Phytochemical investigation12
evaluated for their Antioxidant6, Anti-inflammatory7 action while the
fresh juice has shown the Immunomodulatory & DNA protective The aqueous extract and alcoholic extract of Cynodon dactylon L.
activity8. Phytochemical screening carried in the past has shown the were subjected to preliminary phytochemical investigation using
presence of phenols, flavonoids, alkaloids, glycosides, proteins and standard method of analysis.
amino acid in Cynodon dactylon9.
Identification test by TLC12
Ethnomedicinal investigation revealed the use of Cynodon dactylon L.
as first aid in minor injuries in traditional medicine 10. However, Test solution: Dissolved test extract in 1ml of ethanol.
there is no scientific evidence or report on the wound healing
potential of the Cynodon dactylon L. The present study is planned to Solvent system: toluene: propanol (7:3)
evaluate the wound healing potential of Cynodon dactylon L. aqueous The 20 µl test extract solution was approximately applied on a silica
and alcoholic extract.
gel G plate of uniform thickness of 0.2 mm. The plate was developed
MATERIAL AND METHODS in the given mobile phase up to a distance of 8 cm. On exposure to
the iodine vapour, two brownish yellow spot at Rf value of 0.34 and
Drugs and Chemicals
0.86 was observed.
Povidone-iodine ointment (Betadine), ketamine hydrochloride
Preparation of gel
(Ketamine) distilled water, ethanol was used for study. All chemicals
used were of AR grade. Carbopol 940 forms very good consistent transparent gel at low
Plant collection concentration. Carbopol 940 is non toxic and does not cause any
irritation to skin. So, carbopol 940 was selected as a gelling agent.
Fresh herb of Cynodon dactylon were collected from herbal garden of 1% carbopol gel base was prepared by soaking carbopol 940 in hot
SVKM’s NMiMS, Shirpur campus (M.S.) and sent for authentification water overnight. The formulation was made using below following
to Agharkar Research Institute, Pune.
formula shown in the Table 1.
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Asian J Pharm Clin Res, Vol 5, Suppl 3, 2012, 161-164

Table 1: Composition of gel of aqueous and alcoholic extract of treated with 0.2% w/w betadine ointment, the test groups was
Cynodon dactylon treated with aqueous and alcoholic extract gel (10 % w/w and 20 %
w/w) by applying the it every day till the 16th day of wound healing.
Concentration (% w/w)
Sr.No. Ingradients The progressive changes in wound area were monitored using
10% w/w 20% w/w
1 Carbopol 940 200 mg 200 mg vernier caliper. The measurement of wound area on graph paper
2 Extract 2 gm 5 gm was expressed as unit (mm2). Wound contraction was expressed as
3 Methy paraben 0.2ml 0.2 ml percentage reduction of original wound size.
5 Glycerine 0.4 ml 0.4 ml
Incision wound healing model
6 Triethanolamine 0.2 ml 0.2 ml
7 Water q.s. q.s. This study was carried out (Figure: 1b), as described by James O, et
al. 201015. Tensile strength (the force required to open the healing
Experimental animals
skin) was used to measure the completeness of healing. Tensile
Male wistar albino rats, weighing 200-300 g, were used and housed strength of wound represents the effectiveness of wound healing.
in standard environmental conditions i.e. room temperature 22°C ± Usually wound-healing agents promote the gaining of tensile
3°C, relative humidity 50-60%. The experimental protocol was strength.
approved by the Institutional Animal Ethics Committee (IAEC) of The grouping of animal was done in the same manner as excision
NMIMS, SPTM, Dhule, Maharashtra and conducted according to the model. Six groups of animals containing six in each group were
guidelines of the Committee for the Purpose of Control and
taken. The animals were anaesthetized by ketamine injection. One
Supervision of Experiments on Animals (CPCSEA).
full thickness paravertebral incision of 5 cm length was made
Excision wound healing model including the cutaneous muscles of the depilated back of each rat.
Full septic measures were not taken and no local or systemic
Excision wound healing model (figure: 1a) was performed as per antimicrobials were used throughout the experiment. After the
Mukherjee PK, et al. 200014. This model was used to monitor wound incision was made, the parted skin was kept together and stitched
contraction and epithelialisation time. The animals were divided in with sutures, 1 cm apart. The ointment of the extract, standard drug
six groups having six animals in each group as shown in Table 2. (betadine ointment) was applied to the wound of test & standard
group twice daily, until complete recovery to the respective groups
of animals. On 8th day after wounding the sutures were removed
and the tensile strength was measured on 10th day.
For measuring the tensile strength the rats were again anaesthetised
and each rat was placed on a stack of towels on the middle of the
board. The amount of the towels could be adjusted in such a way so
that the wound was on the same level as the tips of the arms. The
clamps were then carefully clamped on the skin of the opposite
edges of the wound at a distance of 0.5 cm away from the wound.
The longer pieces of the fishing line were placed on the pulley and
finally to the polyethylene bottle. The position of the board was
adjusted so that the bottle receives a rapid and constant rate of
Fig 1: (a) Excision wound and (b) Incision wound model at day 0 water from a large reservoir, until the wound began to open. The
amount of water in the polyethylene bag was weighed and equated
Table No.2: Grouping of animals and their treatment as the tensile strength of the wound. The tensile strength induced by
Group the extract and by betadine ointment treated wounds was compared
Sr. No. Treatment with the control.
(n=6)
1. Control Paraffin wax Evaluation was done by measuring wound area in mm2 in excision
2. Standard Povidone Iodine wound healing model and tensile strength measurement in incision
3. Test -1 Aqueous extract (10%) gel of C.dactylon wound healing model.
4. Test -2 Test aqueous extract (20%) gel C.dactylon
5. Test -3 Test alcoholic extract (10%) gel C.dactylon Statistical evaluation
6. Test -4 Test alcoholic extract (20%) gel
Data obtained are presented as means ± standard error of mean
All the animals were anaesthetized using ketamine hydrochoride. (S.E.M.) for the number of animals in each group (n = 6). The groups
The rats were depilated on the back and a predetermined area of were compared using one-way analysis of variance (ANOVA)
500 sq.mm of full thickness skin was excised in the dorsal inter followed by Dunnett’s test, P<0.05 was considered significant in
scapular region. Rat’s wounds were left undressed to the open excision wound healing model and P<0.01 was considered
environment for control group. The reference standard group was significant in incision wound healing model.
Table No 3: Effect of Cynodon dactylon extract and betadine on wound contraction in excision wound model
Wound area mm2 Mean (± SEM)
Group
0th day 4th day 8th day 10th day 12th day 14th day 16th day
496.4 469.13 417.63 333.13 271.6** 198.36* 170.66*
Control
(± 2.88) (± 3.92) (± 2.75) (± 2.94) (± 2.28) (± 3.57) (± 4.10)
528.26 470.16 310.66** 238.46** 149.43** 89.3** 26.23**
Standard
(± 3.43) (± 2.65) (± 4.39) (± 3.63) (± 4.35) (± 1.26) (± 1.44)
513.26 483.5 333.8** 249.5** 170.46* 102.53** 74.56**
Test aqueous 10%
(± 2.66) (± 3.45) (± 2.45) (± 4.56) (± 4.23) (± 2.63) (± 1.70)
502.6 463.56 315.13** 229.5** 157.4** 89.53* 52.23**
Test aqueous 20%
(± 2.63) (± 3.71) (± 0.99) (± 4.65) (± 3.40) (± 2.18) (± 3.61)
507.26 473.5 323.8** 239.5** 160.46* 96.2** 54.56**
Test alcoholic 10%
(± 2.66) (± 3.45) (± 2.45) (± 4.56) (± 4.23) (± 2.63) (± 1.70)
504.6 463.56 315.13** 229.5* 154.4** 92.53** 41.23**
Test alcoholic 20%
(± 2.63) (± 3.71) (± 0.99) (± 4.65) (± 3.40) (± 2.18) (± 3.61)

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Asian J Pharm Clin Res, Vol 5, Suppl 3, 2012, 161-164

RESULTS Wound healing results analysis

Preliminary phytochemical screening In the study of wound healing potential, the aqueous extract and
alcoholic extract showed prominent increase in the rate of wound
The preliminary screening of aqueous extract and alcoholic extract healing of rats when compared to the control group. In the excision
showed the presence of phenols, alkaloids, glycosides, flavonoids, wound healing model, the test extract gel of Cynodon dactylon L.
tannins, carbohydrate, proteins and amino acid. showed significant wound healing rate when compared to control
group (p < 0.05) as shown in Table 4.
Table No 4: % wound area healed on different days in excision wound model
% wound area healed on different days
Group
0th day 4th day 8th day 10th day 12th day 14th day 16th day
Control 0.00 6.92 32.15 44.97 60.09 72.41 87.52
standard 0.00 8.99 38.83 52.70 69.37 83.86 94.49
Test aqueous 10% 0.00 7.85 34.87 50.95 66.98 79.57 89.37
Test aqueous 20% 0.00 8.25 36.44 52.73 68.24 80.77 90.03
Test alcoholic 10% 0.00 8.13 35.17 53.21 68.73 81.13 91.19
Test alcoholic 20% 0.00 8.87 37.17 54.73 69.59 82.23 92.57
In the incision wound healing model, the aqueous extract & alcoholic
extract of Cynodon dactylon at 10th day was measured and found to ACKNOWLEDGMENTS
be increase the wound healing rate in a manner, and the results
were statistically significant (p < 0.01) as given in Table 5. The The authors wish to thank Mustafa Pata and Ravikumar Sinojia for
wound healing of alcoholic extract was well comparable to the their technical help, and Associate dean with all Management
standard drug. There was less effect with the aqueous extract when members of NMIMS, School of Pharmacy and Technology
compared with alcoholic extract. Managment, Shirpur, for providing the necessary facilities to carry
out the research work.
Table No 5: Tensile strength of different group.
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